PONE-D-23-27932Occurrence and characteristics of extended-spectrum-β-lactamase- and pAmpC-producing Klebsiella pneumoniae isolated from companion animals with urinary tract infectionsPLOS ONE

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This work was supported by a grant (NSTC-112-2313-B-002-031) from the National Science and Technology Council, Taiwan to K.-S. Yeh. The authors extend their gratitude to Dr. Lee-Jene Teng from the Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University, for generously providing the Klebsiella pneumoniae ATCC 700603 strain. Additionally, heartfelt thanks to Dr. Chao-Tsai Liao from the Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, for kindly provision of the Escherichia coli J53 strain.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: N/A

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: 1. The article is well written. The abstract is ok and well-described.

2. The introduction part requires to describe the pathogenic properties of K. pneumoniae a bit stating the important genes and their importance.

3. The materials and methods are well described. A few techniques (like detection of the bacterium by Vitek-2 CMD System) require to be supported by suitable references.

4. Detection of the virulence genes in K. pneumoniae isolates should be discussed in the introduction part too.

5. The results are well written but in complex manner. This part should be properly edited to make the results easily understandable with inclusion of more tables or graphs.

6. The discussion part is well written. All the parameters are well discussed. Comparative analysis with other studies are ok. Few more references supporting the results can be included to support the resistance properties of the ESBL producing K pneumoniae isolates.

7. The conjugational assay and the virulence gene detections are also lacking proper supporting references. Slight elaborate discussion of these parts will make the article more acceptable.

8. The conclusion part is well written.

9. The references are ok and must be in accordance with the journal format.

10. The figures and tables are ok and well described.

Reviewer #2: 1- Line 39, write ‘Klebsiella pneumoniae is a Gram-negative, nonmotile, encapsulated bacterium.’ In all the manuscript write ‘Gram’ not ‘gram’.

2- Line 50, write ‘..ESBL genes are mainly located on plasmids of various molecular sizes, which enhance their horizontal transfer; in addition, those plasmids can confer resistance to other antibiotics such as fluoroquinolones and aminoglycosides [10].’

3- Line 63, write ‘….causing various diseases, such as liver abscess, endophthalmitis,…’

4- Line 169, write: The disc agar diffusion method was used for antimicrobial susceptibility testing [23], and the selected drug discs were as follows: amoxicillin-clavulanic acid (20/10 μg), 171 ampicillin (10 μg), augmentin (5 μg), cefixime (5 μg), cefotaxime (30 μg), cefovecin (30 μg), cefoxitin (30 μg), cefpodoxime (10 μg), cephalothin (30 μg), ciprofloxacin (30 μg), doxycycline (30 μg), enrofloxacin (5 μg), gentamicin (10 μg), imipenem (10 μg), tetracycline (30 μg), and trimethoprim-sulfamethoxazole (25 μg).’

5- In materials and methods section , please indicate the number of isolates that you tested to search the transferability of ESBL/pAmpC transfer. Since it is not clear you tested only one train or many strains, readers realize that you tested 41 isolates only in section results. You can say that only sodium azid resistant isolates were tested for conjugaison; you said this in section results but it is better to say that in section MM.

6- In the beginning of the section discussion you spook about results reported previously in china, Japan (line 337), but you do not provide reference articles of these data, please add the references.

7- In section discussion, generally K. pneumonia contains the SHV-1 (according to literature) so how you explain the absence of this gene in the majority of your strains, provide one or two sentences to explain this.

8- Line 376 you said ‘ST11, accounting for 12.3% (7/57)’ but in line 377 you said ‘.Furthermore, ST11, ST15, and ST655 were detected in 10.5% (6/57) of the isolates each..’ please verify the frequency of ST11 is 12.3% or 10.5%.

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Reviewer #1: No

Reviewer #2: Yes: Mohamed Salah Abbassi

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Occurrence and characteristics of extended-spectrum-β-lactamase- and pAmpC-producing Klebsiella pneumoniae isolated from companion animals with urinary tract infections

PONE-D-23-27932R1

Dear Dr. Yeh,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Nabi Jomehzadeh, Ph.D (Assistant Professor)

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: The article is well revised. All the comments and queries are addressed. The revised article is well explained in all the sections from abstract to conclusion.

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If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

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